PROJECT SUMMARY/ABSTRACT People with HIV-1 (PWH) experience chronic inflammation that leads to comorbidities despite virological suppression on anti-retroviral therapy. These individuals have high rates of substance abuse including cannabinoids which have known immunomodulatory effects. Recent evidence has suggested an important role of the NLRP3 inflammasome as a driver of inflammation. Cannabinoids have been shown to reduce inflammasome activity. What remains unknown is the effect that cannabinoids have on HIV-stimulated NLRP3 inflammasome activity. Our goal is to elucidate the role of cannabinoids in modulating HIV-inflammasome signaling in lymphoid tissue. This study is the proposal of an early stage investigator physician scientist who will bring together expertise in HIV virology, immunology, receptor signaling, and genomics to test the following aims: 1) To determine the effect of cannabinoid treatment on HIV-stimulated NLRP3 inflammasome signaling. 2) To identify the cell types impacted by cannabinoid treatment on HIV-stimulated NLRP3 inflammasome signaling. 3) To identify the impact of cannabinoid treatment on inflammatory signaling in infected and neighboring cells through spatial transcriptomic analysis of infected tonsil blocks. The methods proposed are novel in that they utilize a recently developed ex vivo human tonsil infection model and a single cell imaging and gene expression platform that will allow for understanding the inflammatory signaling that is associated with HIV-1 infection on a single cell level. The investigators and collaborators offer complimentary expertise and the facilities at the Mount Sinai Health System are uniquely capable of supporting these studies with core facilities, large patient cohorts, and institutional support. The Icahn School of Medicine at Mount Sinai has demonstrated commitment to the career development of this early stage principal investigator. The goal is to develop a successful career of the PI as an independent physician scientist in dissecting out the pathogenesis of the cannabinoid signaling in HIV- 1 infection and downstream inflammasome signaling with the goal of finding key therapeutic agents in the treatment of HIV-1 infection and inflammation and in understanding the interplay of chronic inflammation and drugs of abuse.